Low-toxicity pimaricin derivative as well as preparation method and application thereof
A technology of pimaricin and its derivatives, which is applied in the development and preparation of low-toxic pimaricin derivatives, which can solve the problems of strong cytotoxicity, poor water solubility, and restrictions on the development and application of polyene antibiotics, and reduce the Cytotoxicity, good antifungal activity
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Embodiment 1
[0042] Embodiment one: antifungal activity detection
[0043] A. Use Candida albicans SC5314 (ATCC MYA-2876) as the experimental standard strain. Candida albicans clinical strains (CCC-593, CCC-495, CCC-487, CCC-515, CCC-496, CCC-2260, CCC-2251, CCC-2242, CCC-2233, CCC-2224) were used as indicator bacteria.
[0044] B. Select a single colony of Candida albicans to prepare a bacterial suspension in the medium, and the final concentration of the bacterial solution is about 0.5-2.5×10 4 CFU / mL. Take 200 μL of the bacterial solution and add it to the first well of a flat-bottomed 96-well plate, and then add 100 μL of the bacterial solution to be tested to each well.
[0045] C. Add the drug to be tested or the positive control (amphotericin B) into the first well, pipette and mix in the first well, take 100 μL of the uniform suspension and add it to the second well, pipette and mix, repeat to the last well, Perform a 2-fold serial dilution and discard the excess bacterial solutio...
Embodiment 2
[0048] Example 2: Cytotoxicity detection
[0049] A. Oral epithelial cells are cultured in complete medium in vitro. After the cells adhere to the wall and grow to the full, digest and collect the cells, and resuspend the cells in the complete medium.
[0050] B. Each well of the 96-well plate was inoculated with 5×10 3 100 μL of cell suspension. Cell culture incubator (37°C, 5% CO 2 ) culture, and the cells were replaced with serum-free DMEM medium overnight when about 80% of the cells adhered to the wall.
[0051] C. Gradient half-dilution to obtain different concentrations of the drug to be tested or amphotericin B standard solution, add serum-free DMEM medium respectively, and the treatment time is 24 hours.
[0052] D. According to the instructions of the commercially available kit, CCK8 experiment was carried out to detect the inhibitory rate of the compound on oral epithelial cells.
[0053] E. There are 3 parallel experiments for each group of experiments, and each...
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